Systems Integration of a Surface Acoustic Wave Biosensor for Point-Of-Care-Diagnostic Use

The present application is related to and claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Patent Application No. 63/494,315, entitled “SYSTEMS INTEGRATION OF A SURFACE ACOUSTIC WAVE BIOSENSOR FOR POINT-OF-CARE-DIAGNOSTIC USE,” filed Apr. 5, 2023. The entire contents of the aforementioned patent application is incorporated herein by this reference.

The present disclosure relates to an apparatus and methods for identifying chemicals, toxins, gaseous compounds and/or biomarkers related to disease or wellness of humans or animals, as well as environmental issues including for instance, infectious disease (e.g., bacterial, fungal, parasitic infections, viral infections, etc.), chemotoxins, biotoxins and biomarkers (e.g., Glial Acidic fibrillary protein (GFAP), Ubiquitin carboxy terminal hydrolase (UCL1), troponin, and the like). More particularly, the disclosure relates to integrated (both surface and bulk) acoustic wave sensor systems for detecting biomarkers such as protein or nucleic acids, as well as infectious agents such as bacteria or viruses and whole or parts of eukaryotic cells such as cancer cells. BACKGROUND OF THE DISCLOSURE It is important to detect and analyze infectious disease, biomarkers, etc. of interest to effectively treat patients and prevent severe disease as rapidly as possible at the point of care without loss of crucial time. Currently available diagnostic systems utilize either optical (light), molecular, or electrochemical methods of detection and analysis. Disadvantageously, these systems lack portability and require complex and costly instruments. A further disadvantage of these systems is that they require the use of expensive consumables (e.g., multiple reagents), which require extensive sample processing, and result in low signal sensitivity and specificity. There is an urgent and unmet need for an efficient and accurate point of care diagnostic system that facilitates rapid detection of infectious diseases, biological markers (e.g., amino acids, nucleic acids, and the like), or environmental markers (e.g., gaseous, chemical, liquid, and the like) of interest that are more accurate, portable and rapid and applicable to human, animal, and environmental diagnostics. The detection of biological and chemical agents needs to be conducted in biological material such as plasma, blood, nasal swabs, etc. but also in other mediums such as waste water, potable water, water and liquids obtained from contaminated or non-contaminated environmental sources. Such chemical analytes can include a series of chemicals found in human blood and other fluids, including ions such as sodium, calcium, potassium, and enzymes. Other chemical agents such as Alkaline phosphatase, Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Bilirubin (total and direct), Blood glucose, Blood urea nitrogen, Calcium (Ca) in blood, Carbon dioxide (bicarbonate), Chloride (CI), Cholesterol and triglycerides, Creatinine and creatinine Clearance, Gamma-glutamyl transferase (GGT), Lactate dehydrogenase Phosphate in blood, Potassium (K) in blood, Sodium (Na) in blood, Total serum protein, and Uric acid in blood may also be used.

The present disclosure describes an integrated and mutually dependent system and method for diagnosing infectious disease, such as bacterial, fungal, parasitic infections, viral infections, and infectious disease caused by viruses, such as SARS-COV-2, for example, and many non-infectious biomarkers such as hormones, proteins, nucleotides whole or part of cells etc. amongst many others of biological interest, including determining real time biological binding activity such as real time binding of affinity agents such as antigen antibody binding dynamics in relevant biological materials ex vivo. Source of biological agents could include biological samples and other sources such as waste water, potable water and ground waters. The disclosure also includes utilization of the disclosed sensor for a variety of chemical toxins such as PFAS (per- and polyfluoroalkyl substances) to be detected in a variety of water sources such as waste water, ground water and other carrier samples such as food, devices among other sources, Sources of biologically important chemical substances found in blood can include the following: Alkaline phosphatase, Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Bilirubin (total and direct), Blood glucose, Blood urea nitrogen, Calcium (Ca) in blood, Carbon dioxide (bicarbonate), Chloride (Cl), Cholesterol and triglycerides, Creatinine and creatinine Clearance, Gamma-glutamyl transferase (GGT), Lactate dehydrogenase Phosphate in blood, Potassium (K) in blood, Sodium (Na) in blood, Total serum protein, Uric acid in blood among others. The disclosed system employs integrated surface acoustic wave sensor technology (also defined previously) in an efficient, low-cost integrated surface acoustic wave (SAW) biosensor based system for point-of-care diagnostic use that is able to reliably identify biological samples having specific infectious agents and/or biomarkers along with an enhanced detection system and the integrated connectors and software to activate such a system and to provide the analytical tools and user interface for accurate biodetection. Small sample volumes not requiring any or much separation techniques enhance the use of these systems at the point of care and remote situations for humans and animals in both resource rich and resource poor settings.

Aspects of the present disclosure include an integrated surface acoustic wave biosensor in a uniplex or multiplex format system for point-of-care diagnostic along with a preferred metallization waveguide (e.g., delay line) and or a different metallization of other elements of the sensor such as interdigitated transducer (IDT) and reflectors, using various metals and a liquid well containing cartridge adapted specifically for this system. The integrated surface acoustic wave biosensor may produce surface or bulk acoustic waves. The system includes a disposable cartridge component and a reusable reader enclosing a specific contact region whereby data is transferred with integrity from the sensor. It also includes the workings of a reusable reader and ability to multiplex the system for multiple uses and multiple sources of biological materials. According to aspects of the present disclosure, the disposable cartridge component includes a sample well (of open or closed configuration) for addition of the biological sample, integrated surface acoustic wave (SAW) biosensor(s), Radio frequency (RF) and pogo pin and other similar connectors to the SAW biosensor, overmold components and gasket(s). The disposable cartridge also includes a cassette for housing the sample well, the SAW biosensor, the RF and pogo pin and other connectors, and overmolds and gasket(s). Out of cartridge systems for sample processing may also be included, as are reagent tubes that contain a number of reagents such as gold nanoparticles and buffers. According to aspects of the present disclosure, the reusable reader includes a RF port(s) to connect to the RF connector(s) that is (are) part of the disposable cartridge component. The RF connector(s) interface with the SAW biosensor(s) via pogo pins or other such direct connectors that are integrated with the disposable cartridge component. The biosensor may be secured to the cartridge via a galvanic connection assembly.

The disposable cartridge may also include an absorbent wicking pad which allows for repeat of fluid allocation and a simple washout. In illustrative embodiments, the disposable cartridge is attached to the sensor utilizing an overmolded gasket or a felt seal. The advantage of the felt seal to cover the IDTs may be preferential as a way to control fluids on the SAW biosensor as it does not apply significant pressure and dampening to the surface. However, any appropriate sealing device (e.g., a gasket, a felt seal, an overmold, a fabricated wall, or the like) will be acceptable. A fabricated wall may be comprised of a silicone rubber material adhered along the perimeter of a sensing area of the integrated acoustic wave biosensor. In another illustrative embodiment, the SAW biosensor includes a sample channel and one or more reference channels, optionally coated with at least one metal. In another embodiment, the biosensor may include at least two contact pads. In another embodiment, the sensor can be multiplexed with multiple waveguides on a single sensor platform and an adapted flow cell to flow over the entire multiplexed system. In another illustrative embodiment, the SAW biosensor can be multiplexed with multiple uniplex sensors in an array and an adapted flow cell to flow over the entire multiplexed system. In another illustrative embodiment, two or more uniplex and/or multiplex SAW biosensors that each have their own open sample well can be combined into one unit for a multi-well cartridge. In another illustrative embodiment, the SAW biosensor includes a piezoelectric crystal base, such as a lithium tantalite crystal base. Another aspect of the present disclosure provides a disposable cartridge component of an integrated surface acoustic wave biosensor system for point-of-care diagnostic use. The integrated acoustic wave biosensor may also include a coded reflector system comprised of one or more metals. The integrated acoustic wave biosensor may also include uniplex or multiplex sensors which generate surface acoustic waves in the range of 50-1000 MHz or bulk acoustic waves in the range of 50-5,000 MHz.

In some instances, the sample added to the well may need to be mixed in order to make sure that the analytes can adequately interact with the SAW biosensor surface. In this case, we propose a closely held but external vibration module which can mix the fluid in an open well cartridge flow cell as needed.

The biological sample may require processing prior to its addition to the sample well on the disposable cartridge. Materials for off cartridge sample preparation includes, but is not limited to, reagent tubes containing buffer solutions, reagents, such as gold nanoparticles, which may be lyophilized or in solution, syringes, and syringe filters.

Another aspect of the present disclosure provides a method for detecting a target analyte in a biological sample using an integrated surface acoustic wave biosensor system. According to aspects of the present disclosure, the method includes steps of providing a disposable cartridge component of the integrated surface acoustic wave biosensor system, providing the biological sample into a sample well of the disposable cartridge component to interact with the propagating SAW to generate a characteristic electrical signal of the biological sample, and detecting the target analyte based on the characteristic electrical signal.

In one aspect, the disclosure provides an integrated acoustic wave biosensor system, which includes, a disposable cartridge component, which includes, a cassette including: a radiofrequency (RF) port configured to receive an RF signal; a well for the addition of a sample in a fluid; an integrated acoustic wave biosensor, wherein the integrated acoustic wave biosensor includes: a piezoelectric base; at least one metal coated channel; a coded reflector system; at least one sensor, at least one set of interdigital transducers (IDTs), and at least two contact pads; a galvanic connection assembly coupled to the integrated biosensor; and a sealing device configured to separate the fluid from the integrated acoustic wave biosensor; wherein, the integrated acoustic wave biosensor is activated an RF source.

In some embodiments, the sealing device is a fabricated wall.

In some embodiments, the fabricated wall is comprised of a silicone rubber material adhered along a perimeter of a sensing area of the integrated acoustic wave biosensor.

In some embodiments, the sealing device is a polymer gasket or a felt seal.

1 In some embodiments, a layer of PMMA, silane, or silicone dioxide isolates the region comprising the IDTs, one or more waveguides, and a coded reflector system from the fluid and allows for a rubber gasket around the perimeter of the region comprising the IDTs, waveguides, and reflectors of the integrated acoustic wave biosensor.6. The disposable cartridge component of claim, wherein the disposable cartridge is configured to be paired with a cartridge reader In some embodiments, the integrated acoustic wave biosensor is a surface wave acoustic biosensor or a bulk wave acoustic biosensor.

In some embodiments, the well leads to a fluid flow over the sensor in a single plex or multiplex format.

In some embodiments, the well is an open well.

In some embodiments, the well is a closed well.

In some embodiments, the coded reflector system is of (HFM) or (OFM) configuration.

In some embodiments, the disposable cartridge further comprises two or three channel sensors or one or more multiplex sensor(s).

In some embodiments, the coded reflector system comprises one or more metals.

In some embodiments, the one or more metals are selected from the group consisting of gold or aluminum.

In some embodiments, unidirectional IDTs are used instead of the coded reflector system.

In some embodiments, the disposable cartridge comprises a felt seal to protect the IDTs and the coded reflector system.

In some embodiments, the one or more sensors are uniplex sensors or multiplex sensors.

In some embodiments, the uniplex or multiplex sensors generate surface acoustic waves in the range of 50-1000 MHz or bulk acoustic waves in the range of 50-5,000 MHz.

In some embodiments, the surface acoustic waves are in the range of 100-600 MHz.

In some embodiments, the surface acoustic waves are in the range of 100-900 MHz.

In some embodiments, the disposable well cartridge system has a wicking pad.

In some embodiments, the disposable well is closed.

In some embodiments, the disposable well is open.

In some embodiments, the gasket is made of polymer or overmolded TPE onto the well.

In some embodiments, the polymer is PDMS, other silicone-based materials or other low durometer polymer materials.

In some embodiments, the biosensor comprises a sample channel and a reference channel and optionally comprising a compensation channel.

In some embodiments, the acoustic wave biosensor has a metallic waveguide comprising one or more metals selected from the group consisting of gold, titanium, chromium, and aluminum.

In another aspect, the disclosure provides a disposable cartridge component of an integrated acoustic wave biosensor system, which includes: a well for the addition of a fluid sample; a cassette comprising an integrated acoustic wave biosensor, a galvanic connection assembly coupled to the integrated acoustic wave biosensor, and a sealing device separating the sensing area from the integrated acoustic wave biosensor; and a docking location on the cassette body configured to couple with a well.

1 In another aspect, the disclosure provides a method for detecting a target analyte in a biological sample using an integrated acoustic wave biosensor system of claim, the method comprising: providing the fluid sample into the well; applying surface acoustic waves to the sample in the well to generate a characteristic electrical signal of the biological sample; and detecting the target analyte based on the characteristic electrical signal.

The present disclosure is based, at least in part, on the discovery that a gold overlay (to attach biological agents) and coded reflector integrated acoustic detection device such as a Surface Acoustic Wave (SAW) device with an optional dual metallization of gold overlay on the waveguide and a different metal such as aluminum on the interdigitated transducers (IDTs) and reflectors for reduction of interdigitated reflection can provide extremely sensitive detection of biomarkers and infectious disease-related antigens (e.g., SARS-COV-2, SARS-COV, MERS-COV, human coronavirus OC43, human coronavirus HKU1, human coronavirus 229E, human coronavirus NL63 and the like) in a sample. Infections afflicting animals, such as canine heart worm, equine viruses and feline viruses, can also be detected. Other examples include antigen driven circulating antibodies, protein biomarkers of interest in wellness and disease and cancer markers on cells and in liquid phase.

Aspects of the present disclosure include platforms, apparatus, and systems that are useful for the identification of biological agents, chemicals, toxins, environmental agents and the like. The disclosed platforms, apparatus, and systems may be used for diagnosis, treatment and/or prevention of any biological event of interest such as cardiac events, neurological events reproductive events, cancer, etc., as well as a variety of infectious diseases such as, e.g., those caused by bacteria, immunological events, fungi, viruses, eukaryotic cells, proteins, nucleotides, and the like. The use of this biosensor is disclosed for human, animal (e.g., companion, wildlife, and food animals), and environmental use. The disclosed apparatus, systems and platforms may be used for detection of non-biological systems, such as chemotoxins and gaseous systems.

The use of acoustic based analysis systems for biosensing (e.g., general methods of acoustic analysis, reflector-based and unidirectional IDT-based surface acoustic waves, bulk acoustic waves, and the like) has previously been proposed but not implemented in various biosensor designs for use in point of care diagnostic systems and detectors. In particular, the techniques herein provide an acoustic wave sensor known as a Surface Acoustic Wave (SAW) sensors, which operate on the principle of passive wireless sensing of a mass/viscosity change using piezoelectricity as a sensing platform, that allows electronic detection of specific parameters when changes to the acoustic wave properties are conveyed as a change in electronic measurements. Piezoelectricity is a phenomenon displayed in certain crystals, such as quartz, lithium niobate, and lithium tantalate, where voltage generation is induced by mechanical stress. Interestingly, the reverse is also true, such that application of voltage will induce a mechanical deformation or stress. The ability of the piezoelectric crystals to undergo atomic vibration in the presence of an electrically generated radio frequency input presents the ability of crystals to function as sensors. SAW sensors are used in the detection of changes in mass, viscoelasticity, conductivity, and dielectric properties derived from mechanical or electrical variations. SAW sensors also employ the piezoelectric effect to excite acoustic waves electrically at an input transducer and to receive the waves at the output transducer.

Previous publications of this applicant have described the use of different coatings, layers, signal amplification, interfaces with fluid materials, connectors of various sorts, cartridges and fluid control systems and multiplexing for use in various sizes of SAW-based biosensors. Examples of such publications include U.S. patent application Ser. No. 16/629,309 filed Jan. 7, 2020, entitled “BIOACTIVE COATING FOR SURFACE ACOUSTIC WAVE SENSOR”; U.S. patent application Ser. No. 16/629,305 filed Jan. 7, 2020, entitled “METHODS AND APPARATUS FOR INTERFACING SENSORS WITH FLUID MATERIALS”; U.S. patent application Ser. No. 16/629,307 filed Jan. 7, 2020, entitled “MULTIPLEXING SURFACE ACOUSTIC WAVE SENSORS WITH DELAY LINE CODING” the disclosures which are hereby incorporated by reference in their entirety.

Piezoelectric crystals, such as quartz, lithium niobate and tantalate, that support surface type acoustic waves are typically only weakly responsive to adequate adhesion of biological materials. Chemical agents such as silane compounds following a series of proprietary application procedures along with reactive functional groups, such as amine residues, have been used to enhance adhesion of biological molecules on the surface crystals.

The disclosure provides techniques for modifying the crystal surface of SAW based biodetectors by applying a layer of gold, silica or aluminum onto the surface of the crystal during fabrication which also serves as a waveguide in directing the acoustic wave in the right orientation in our configuration of a reflected wave. This approach in which the metalized waveguide at least partially coats the surface of the crystal with a metal is more amenable to the attachment of biological molecules. Additionally, it is contemplated within the scope of the disclosure that another metal can be used to construct the IDTs and reflectors. Adding a metal layer such as gold on the crystal surface of a SAW biodetector to serve as a critical waveguide has also been attempted. However, while some metal surfaces are more effective on the surface of the propagating acoustic waves, metals such as aluminum bind biological molecules poorly when placed on the waveguide and are therefore not optimal for use on the surface of SAW detection and diagnostic devices. Biological buffers can etch the aluminum waveguide surfaces, which can adversely affect the propagation of the SAW. The present disclosure provides a preferred metal structure consisting of a gold layer on the reflector-based sensor design and flow cell, optionally including another metal on the IDTs and reflectors more amenable to wave generation, which allows the acoustic wave to propagate effectively and is also significantly compatible with biological capture agents binding to its surface via thiol groups. The gold layer thickness is optimized to reduce the signal-to-noise, which enhances the sensitivity of the SAW. Also demonstrated is early data on the sensitivities of binding and detection of proteins at very low concentrations, a major advantage to our system.

Other approaches to modifying the crystal surface of SAW based biodetectors included applying a layer composed of a dielectric material such as SiO2, Poly (methyl methacrylate), or gold, on the surface of the SAW sensor to further trap the energy of the acoustic wave closer to the surface for enhanced analyte detection. Similarly, a layer of silicon dioxide may enhance the binding ability of biological molecules without interfering with the transmission of the surface wave.

A major disadvantage of previously described SAW based detection and diagnostic systems is due, at least in part, to their inability to effectively bind captured agents onto the surface of acoustically transmissive materials in a liquid environment and to carefully separate the liquid environment from the electronic components. The techniques herein overcome these issues, which provides a fully functional point of care diagnostic system for use in human, animal and research based diagnostic systems.

There remains an urgent world-wide need for a rapid, cost-effective, rapid, portable, sensitive, and robust point of care diagnostic test that can be used for the detection of a variety of biological analytes. In this way, the present disclosure fulfills all of these criteria and furthermore, takes advantage of many recent advances in semi-conductor industry (miniaturization, FPGA, software and hardware advances) and advances in cellular communications (on which these sensors are based) to provide a cost effective and easily used system that can replace 60-year-old technologies such as lateral flow test.

Aspects of the present disclosure address this urgent need and overcome the various disadvantages of previously known systems and methods by providing an efficient, low-cost integrated surface acoustic wave (SAW) biosensor based system and method for point-of-care diagnostic use that is able to reliably identify biological samples having specific agents of interest, both infectious and non-infectious.

In one embodiment of the instant disclosure, an integrated surface biosensor system for the use as a rapid, cost-effective, and robust POC diagnostic for the detection of infectious events, biological agents, and systems is provided.

The techniques herein provide acoustic wave-based POC devices suitable for biological events (e.g. infection agent-virus) systems testing. The acoustic devices and methods described herein utilize a piezoelectric material that responds to an electrical signal by generating an acoustic wave (i.e., very high frequency sound) as the fundamental sensing signal.

In addition to this, multiple biochemical analytes in blood or other body fluids can be simultaneously detected on a single sample channel. This paradigm enables multiplex detection on a uniplex SAW biosensor. For this concept, a cocktail of capture agents that can detect the different biochemical analytes of interest (for example Analytes A, B, C) is coated on the surface of the sample channel. The individual SAW changes in the response time and frequency shift of Analytes A, B, and C need to be known from previous experimentation. The unique relationship between the response time and frequency shift for each analyte is used to deconvolute the identification of Analytes A, B and C when binding to the cocktail of capture agents functionalized to the sample channel. This method assumes that binding of one biochemical does not affect the binding of another.

Aspects of the present disclosure include a disposable cartridge system utilizing a surface acoustic wave (SAW) biosensor with a metallized waveguide of various metals as described that can be used for the detection of biological agents and determine biological binding effects in real time. The cartridge can also include an absorbent wicking pad made of cellulose or the like for washout and repeat of fluid allocation. In addition, a vibration module that is external to the cartridge can provide a method to mix reagents in the flow cell. Fluidic sealing is accomplished using polydimethylsiloxane (PDMS) or the like, thermoplastic elastomers (TPE), or felt seals.

1 FIG.A 100 116 112 114 100 106 108 108 100 102 104 100 102 100 presents an exemplary design of a 300 MHz (50-999 MHz) SAW biosensor. Although this exemplary embodiment describes a 300 MHz sensor, it is to be understood that the surface acoustic waves disclosed herein can have various frequencies ranging from 50-1000 MHz (+/−25 MHz). For example, the frequency of a surface acoustic wave can be about 50, about 100,about 150, about 200, about 250, about 300, about 350, about 400, about 500, about 600, about 700, about 800, about 900, or about 1000 MHz (+/−approximately 25 MHz). Similarly, the frequency of the surface acoustic waves can be less than 100 MHz, greater than 100 MHz, greater than 300 MHz, greater than 500 MHz, or greater than 1000 MHz. The SAW biosensor may be activated by high frequency radiofrequency (RF) waves from a RF source. A sample channel reflectorand reference channel reflectorreturns this wave and this is a coded reflector with a hyperbolic frequency modulation (HFM) or orthogonal frequency modulation (OFM). The SAW biosensorcomprises a piezoelectric crystal baseand one or more metal surfaces. In some embodiments, the one or more metal surfacesmay be coated with a biofilm. The SAW biosensormay include a sample channeland a reference channel. A biological sample may be contacted with the SAW biosensorvia the sample channelby dispensing the biological sample with a pipette or dropper into a cartridge flow cell enclosing the SAW biosensoronto which the biological sample may be analyzed.

102 118 119 106 In some embodiments, sample channelmay be coated with one or more capture agents. In further embodiments, the sample channel sensing areaand the reference channel areaof the SAW device may be a metalized (e.g., Al and/or Au layer) and the piezoelectric crystal basechosen may be, e.g., Lithium tantalate (LiTaO3), lithium niobate (LiNbO3), and quartz. The SAW generated on this crystal may be called a Leaky wave, which may be principally composed of a shear horizontal wave so it can operate in a liquid while keeping a low propagation loss. In some embodiments, a third channel (not shown in the figure) may be added to the SAW biosensor, which serves as a second reference channel to remove the effect of ambient temperature change on the SAW biosensor.

1 FIG.B 120 123 122 120 128 120 120 102 120 125 104 102 125 104 122 123 130 132 123 122 102 125 104 102 125 104 123 122 shows an exemplary design of a 50-999 MHz MHz 3-channel SAW biosensor, which has been modified with larger metal contact pads and a third channel for temperature compensation. While the metallization of all the components could be made of gold, it is also possible that the IDTsand reflectorcould be made of a different metal. The disclosed 3-channel SAW biosensorincludes 3-channel contact padsfor establishing a connection between the 3-channel SAW biosensorand a printed circuit board (PCB) (not pictured) supporting it. The disclosed 3-channel SAW biosensorincludes a sample channelfor contacting the biological sample with the 3-channel SAW biosensor, a compensation channel, and a reference channel. The sample channel, compensation channel, and reference channeleach have reflectorsand interdigitated transducers (IDTs)for conversion of electrical energy to mechanical energy and vice versa. Probing structure 1and probing structure 2may be used to test the sensors for quality control after fabrication. IDTs, reflectors, and sample channel, compensation channel, and reference channelcould be made of the same metal or different metals can be used for sample channel, compensation channel, and reference channelthat are biocompatible, such as gold, while a different metal, such as aluminum, could be used for electrical transmission on the IDTsand reflectors.

1 FIG.C 1 FIG.A 134 102 110 112 100 shows a side view of the long axisthrough the center of the sample channel, contact pad, and sample channel reflectorof SAW biosensorfor the exemplary design in.

1 FIG.D 1 FIG.A 140 102 104 100 shows a side view of the short axisperpendicular to the sample channeland the reference channelof SAW biosensorfor the exemplary design in.

1 FIG.E 1 FIG.B 150 112 112 130 120 shows a side view of the long axisthrough the center of the sample channel, sample channel reflector, and probing structure 1of 3-channel SAW biosensorfor the exemplary design in.

1 FIG.F 1 FIG.B 160 102 125 104 120 shows a side view of the short axisperpendicular to sample channel, compensation channel, and reference channelon 3-channel SAW biosensorfor the exemplary design in.

2 FIG. 200 204 100 204 204 208 112 114 100 Referring now to, a schematic comparisonbetween a miniaturized SAW biosensordesign and the larger SAW biosensordesign is presented (frequency range from 50-999 MHz). According to an aspect of the present disclosure, the miniaturized SAW sensormay be designed with a pair of contact pads opposing each other. Miniaturized SAW biosensorhas a miniaturized SAW biosensor reflectorthat is similar to that of the sample channel reflectorand reference channel reflectorin the larger SAW biosensor design.

3 FIG. 3 FIG. 300 300 312 314 316 318 319 300 302 304 306 308 310 300 320 322 324 326 328 402 330 332 334 336 338 312 319 320 338 shows an exemplary design of a multiplexed SAW biosensordesign with multiple waveguides on a single sensor and a gold metal overlay. This disclosed multiplexed SAW biosensorhas five channels: a first channel, a second channel, a third channel, a fourth channel, and a fifth channel; four of which may be used as sample channels and one of which may be used as a reference channel. Each channel has reflectors and IDTs for conversion of the electrical energy to mechanical energy and vice versa. This disclosed multiplexed SAW biosensorhas five reflectors: a first reflector, a second reflector, a third reflector, a fourth reflector, and a fifth reflector. The disclosed multiplexed SAW biosensorhas five sets of IDTs: a first set of IDTs, a second set of IDTs, a third set of IDTs, a fourth set of IDTs, and a fifth set of IDTs. A RF connectormay interface with a first set of metal contact pads, a second set of metal contact pads, a third set of metal contact pads, a fourth set of metal contact pads, and a fifth set of metal contact padsvia pogo pins or other such direct connections (not shown in) that are integrated with the disposable cartridge component for interrogation by the reader. As described above,-may be constructed of a biocompatible metal such as gold while-are constructed of a different more conductive metal such as aluminum for illustration.

4 8 FIG.- Aspects of the present disclosure further includes a disposable open well () cartridge system utilizing a SAW biosensor, RF and pogo pin connection assembly, fluid gasket, and open well frame that can be used for the detection of infectious agents.

4 FIG. 1 FIG.A 400 402 404 100 402 110 404 presents an image of an exemplary prototype connection assemblyof RF connectorand a pogo pinconnector connecting one pogo pin per each channel and a ground pad to the SAW biosensor. A RF connectormay interface with metal contact pads(shown in) via pogo pinsor other such direct connections that are integrated with the disposable cartridge component for interrogation by the reader.

5 FIG.A 500 100 504 502 504 502 110 123 122 510 100 512 Referring now to, an exemplary schematic of a fabricated wallfor a SAW biosensor is presented. To ensure that the IDTs and reflectors, which drive the SAW biosensor, may be protected from the fluid flow of saline and the biological sample, a fabricated wall concept may be adopted to eliminate the need for a gasket. A leaking fluid wall may attenuate the SAW signal conveying the radiofrequency wave into the crystal via the metal contact pads and IDTs leading to signal loss, affecting the functioning of the sensor. In one embodiment of the present disclosure, a fabricated wall concept may be adopted where the SAW biosensorhas been identified to have open sensing areasand liquid-proof protecting areas. Open sensing areasmay be identified on the sensor via fiducials. In the liquid-proof protecting areas, regions of contacts pads, regions of IDTsand regions of reflectorsare isolated from the fluid flow via lid silicon glassadhered on to the SAW biosensorvia seal-proof silicon rubber wall. Alternatively, liquid-proof protection may be achieved by depositing a layer of polymethyl methacrylate (PMMA) or silicon dioxide on the SAW sensor.

5 FIG.B 520 530 538 100 530 110 123 122 532 530 504 depicts a side viewwhere a layer of PMMA, silane or silicon dioxideis overlayed on top of the metal structuresof the SAW biosensor. The layer of PMMA, silane, or silicon dioxideisolates the entire SAW sensor surface from the fluid flow, including the regions of contacts pads, regions of IDTsand regions of reflectors. An additional biocompatible metal layer, such as gold, can be deposited on top of the layer of PMMA, silane, or silicon dioxidein the open sensing areasfor the immobilization of capture agents to the SAW sensor.

5 FIG.C 530 538 502 100 560 502 110 123 122 depicts another such embodiment, where PMMA, silane, or silicon dioxideis overlayed on top of the metal structuresin the regions of liquid-proof protecting areasof the SAW biosensor. The layer of PMMA or silicon dioxideisolates the regions of liquid-proof protecting areasfrom the fluid flow, including the regions of contacts pads, regions of IDTsand regions of reflectors.

6 FIG. 6 FIG. 8 FIGS.A-B 13 FIG.B 15 FIG.B 600 600 600 604 Referring now to, an exemplary diagram of a polydimethylsiloxane (PDMS) or some such polymer gasketis presented. To further isolate fluid flow from the sensitive electrical components, the system may include a polymer gasketto prevent the SAW biosensor from directly interacting with the fluid flow. The polymer gasketcomprises a polymer fluidic channelcreated by the gasket to allow the biological sample to flow through. In some embodiments, as in the case of, the gasket may be made of PDMS or other low durometer polymer material. Disposable cartridge systems in,, andmay incorporate this polymer gasket.

7 FIG.A 700 700 704 702 706 Referring now to, an exemplary SAW biosensor systemwith an enclosure comprising overmolded thermoplastic elastomers (TPE) is presented. The SAW biosensor systemmay include an overmolded TPE gasketon a top surfaceof a cassette component. The system may be completely enclosed using walls of silicone or overmolded thermoplastic elastomers (TPE) which are precisely tooled to the dimensions of the sensor, allowing for the characteristics needed for a closed or open well. Such an enclosure would work for acoustic sensors using other types of acoustic waves, such as Rayleigh, shear horizontal SAW with or without an energy trapping layer, transmission or reflective delay line modes, and with bulk acoustic waves. Any type of acoustic wave traversing a piezoelectric crystal could be used in such an enclosure. All electronic components are protected, and the flow cell ties perfectly to the larger fluidic channel.

7 FIG.B 720 724 730 726 724 728 724 depicts the side view of a felt sealas an alternative to the overmolded TPE and PDMS gasket used provide protection from fluid for the SAW IDT and reflector elements. Typically, felts absorb fluids. This is true regardless of the material from which the felt is made because the high surface area of the intertwined small felt fibers interacts with the surface tension of almost any fluid to quickly absorb that fluid into the felt. But this method uses fluid repellent coatings, such as siloxane or perfluorobutanesulfonic acid (PFBS), to apply a very thin water-repellent coating to the patterned felt. The treated feltis then pressed onto the surface of the piezoelectric surfacewith some moderate pressure. Felt fluidic channelscut into the feltthen keep the water in place without allowing the water to migrate from its intended holding spaces or flow channels. The fibersof the felt extend above and below the felt by fractions of a millimeter, and each fiber is hydrophobic, preventing water from leaking above or below the flat feltpiece.

720 728 724 728 724 The other feature of this felt sealis that the flexibility of the individual fibersof the feltthat are touching the biosensor surface are not stiff enough to couple energy from the SAW. While the SAW is typically in the frequency range of tens to hundreds of megacycles per second (megahertz, MHz), the maximum vibration frequency of these fibersis typically measured in tens of kilocycles per second (kilohertz, kHz), several orders of magnitude lower in frequency. Thus, even when the feltis put under significant pressure on the top of the SAW biosensor, no mechanical energy is lost as the MHz mechanical waves propagate across the surface or in the bulk of the device. As these acoustic oscillations are the key to many different methods of chemical and biological sensing, the ability of this invention to hold the water in place without dampening or otherwise disturbing the acoustic waves is an important technological innovation in sensing devices.

724 724 722 724 726 724 The feltpieces are placed in alignment on top of the SAW biosensor and pressed into place either with tape over the feltor by being pressed by a stiff pressing itemsuch as a plastic cover that shares the 2D channel geometry instantiated by the felt. This larger assembly which contains the feltis then ready for addition of liquids into the fluidic channels. While the feltis generally used in planar configurations, the flexibility of the felt allows it to be used in cylindrical fluidic applications as well, where the felt flexibility may allow for good sealing over curved surfaces.

724 728 The density and fiber diameter and other parameters of the feltare chosen to reduce mechanical coupling in the frequencies of interest and also to enable good sealing. Thickness ranges of felt range from 0.1 mm to 5 mm thick, and felt fiberdiameters range from 10 microns to 0.5 mm.

Alternatively, materials such as cellulose and nitrocellulose papers can also be used for this purpose after hydrophobic coatings if the coupling of the cellulose fibers is at a low enough frequency and the paper surface is rough enough. Very flat paper (with surface roughness less than +/−5 microns) typically will allow water to seep past the smooth paper/surface interface. The paper must have roughness larger than +/−10 microns on its surface to work. Multiple other materials such as nylon, polytetrafluoroethylene (PTFE), polyester, Kapton (polyamide) and wool can also be used as the felt material.

7 FIG.C 740 726 724 746 Referring now to, a top perspective view of an exemplary design of the felt sealwith felt fluidic channelscut into the treated feltapplied to the SAW biosensor surfaceis presented. The stiff pressing item is not included for visual clarity.

8 8 FIGS.A andB 800 present perspective and exploded views of an exemplary embodiment of a disposable open well cartridge systemcomprising a SAW biosensor.

8 FIG.A 8 FIG.B 4 FIG. 800 802 804 820 822 828 826 100 600 100 600 822 804 804 100 100 800 800 shows an exemplary embodiment of the disposable open well cartridge systemwhich includes a cassetteand open well.shows an exploded view of a disposable open well cartridge systemwherein a cassette bodyand a cassette bottomare secured together to hold the SAW biosensor. The disposable open well cartridge system may be integrated with a connection assembly of RF and pogo pin connectors similar to the embodiment shown into interface with SAW biosensor. The disposable cartridge system may also include a polymer gasketto separate the biological sample from coming in contact with the SAW biosensor. Polymer gasketintegrates with the cassette bodyto form the open well, which is capable of handling a variety of bodily fluid and reagents. If necessary, the disposable open well cartridge system may also include an absorbent wicking pad (now shown). Following calibration, the biological sample is simply added directly to the open welland may remain there for the duration of the test. The SAW biosensoris in direct communication with the connection assembly which processes the output response of the SAW biosensorto the biological sample. The disposable open well cartridge systemmay be operatively placed in a reader so that an output of the disposable open well cartridge system may be received by the reader. The output of the disposable cartridge systemmay include a measurement or indication of SARS-COV-2 virus being present in the biological sample, as an example and may consist of any number of biomarkers or cells.

8 FIG.C 860 862 870 804 870 868 870 872 866 870 804 868 804 870 804 shows an exemplary embodimentof an open well cartridge flow cellattached to a wicking apparatusto absorb any fluids which need to be washed out for repeat fluid allocation. Certain testing procedures that require the addition of two fluids to the open wellmay necessitate a washout step to remove fluid from the first addition. This can be realized by having a wicking apparatuswith an absorbent wicking pad. For the washout step, the user lifts the wicking apparatusas indicated by arrowsandto set the wicking apparatusin contact with the open well. The absorbent wicking padthen uptakes the fluid from the open well. The user then lifts the wicking apparatusto its initial state. Upon completion, the open wellis ready for repeat fluid allocation.

8 FIG.D 880 884 882 884 888 884 804 presents the use of a vibration module which can mix the fluid in the open well cartridge flow cell. The vibration moduleis mounted next to the cartridge interfacethat is part of the reusable reader. The vibration modulecan mix the fluid in the open well cartridge flow cell. Power and vibration frequency of the vibration moduleare regulated by the reusable reader. The generated vibrations cause the fluid in the open wellto mix, which may be necessary to promote the reaction of the analytes in the fluid with the functionalized SAW biosensor surface.

9 FIG. 300 900 904 300 Referring now to, a perspective view of a multiplexed SAW biosensorand multiplexed cassette bottomwith multiple waveguides on a single sensor and an adapted flow cellto spread over the entire multiplexed system is presented. This concept allows for the same fluid to simultaneously interact with the multiple waveguides on the multiplexed SAW biosensor.

10 FIG. 1002 1000 1004 1002 presents a perspective view of SAW sensors multiplexed with multiple uniplex sensorsin an array, multiple uniplex cassette bottom, and an adapted multiple uniplex flow cellover the entire multiplexed system. This concept allows for the same fluid to simultaneously interact with the multiple uniplexed sensors.

11 FIG. 300 804 900 presents a perspective view of two multiplexed SAW biosensorswith each having its own open welland cassette bottom. The two multiplex sensors are combined into one unit for a multi-well cartridge. According to an aspect of the present disclosure, an input electrical signal from the reader is traversed through IDTs, a waveguide, and then reflected back via reflectors to the IDTs, where it is reconverted into an electrical signal. Changes in phase and amplitude of the electrical signal are measured by the reader. The phase change data and amplitude change data may then be transformed by appropriate signal algorithms for detecting selected target components of a sample. According to an aspect of the disclosure, multiple IDTs can be configured in a series or in an electrical parallel arrangement.

12 FIG.A 9 FIG. 10 FIG. demonstrates a circuit of the electrical parallel arrangement, as the delay line connects the reader and a cartridge. In certain embodiments utilizing a multiplex approach, multiple waveguides on a single sensor (as exemplified in) or multiple uniplex SAW sensors (as exemplified in) are housed together within a cartridge, along with a switch which selects a waveguide or sensor, respectively, to interrogate.

12 FIG.B demonstrates that upon stimulation, the switch routes an RF electrical signal in the range of 50 to 1000 MHz to the sensor currently being interrogated. To provide the switch with energy and the control signal, the galvanic connection enables the transfer of phase change data and amplitude change data associated with the energy transfer, across the circuit. Then, the system uses a 1 MHz carrier on/off control signal for the switch to enable the phase change data and amplitude change data associated with the energy transfer. Additionally, a third signal is rectified and used as energy supply to the switch and to the microcontroller, decoding the on/off keying to control the switch.

13 13 FIGS.A andB 1300 1300 1303 1313 1312 1313 1312 1320 1300 100 1318 1318 1312 1318 1312 1326 1318 600 100 1318 1318 1324 present perspective and exploded views of an exemplary embodiment of disposable cartridge systemcomprising a SAW biosensor for the detection of infectious diseases. The exemplary embodiment of the disposable cartridge systemmay include a disposable cartridge cassetteincluding a cassette bodyand a disposable cartridge cassette bottomsecured together for holding the sample. The cassette bodyand the disposable cartridge cassette bottommay be secured to each other via vertical posts, for example. The disposable cartridge systemalso includes a SAW biosensorattached to a printed circuit board. The printed circuit boardis then attached to the disposable cartridge cassette bottom. The printed circuit boardmay be secured to the disposable cartridge cassette bottomvia screws (not shown in the figure) through the screw holesalong the perimeter of the printed circuit board, for example. The disposable cartridge system also includes a polymer gasketto separate the biological sample from coming in contact with the SAW biosensorand the printed circuit board. The printed circuit boardincludes a connection regioncomprising printed circuit board contacts (not shown).

1303 1305 1307 1304 1302 1310 1302 1308 1300 According to an aspect of the present disclosure, the cassetteincludes outlines of internal fluidic pathway 1, and fluidic pathway 2. A biological sample may be placed in a sample welland secured with a sample capon the disposable cartridge cassette top. In an illustrative embodiment, the sample capincludes an air vent. The disposable cartridge systemmay be secured onto a reader (not shown) for calibration.

1306 1309 1308 1310 1309 1306 1308 1308 1306 1309 1308 100 1304 Following calibration, a saline capcomprising a springis pushed down on a saline-saturated compression padon the disposable cartridge cassette top. The rebounding spring force of the springin the saline capmay be designed to allow the saline-saturated compression padto decompress and pull the saline back into the saline-saturated compression padat a controlled rate. The saline capis then pushed down against the springcausing the saline in the saline-saturated compression padto flow through a channel over the SAW biosensorto the sample well.

100 1318 100 1300 1300 The SAW biosensoris in direct communication with the printed circuit boardwhich processes the output response of the SAW biosensorto the biological sample. The disposable cartridge systemmay be operatively placed in a reader so that an output of the disposable cartridge system may be received by the reader. The output of the disposable cartridge systemmay include a measurement or indication of SARS-COV-2 virus being present in the biological sample, for example.

14 14 FIGS.A andB 1400 1400 1412 1413 1404 1402 present a perspective and exploded view of another illustrative embodiment of second disposable cartridge systemcomprising a SAW biosensor for the detection of infectious diseases according to aspects of the present disclosure. The second disposable cartridge systemcomprises a second disposable cartridge cassette bodyand a cassette top coversecured together and comprising a SAW biosensor, a printed circuit board, and a gasket to prevent the biological sample from directly being contacted with the SAW biosensor. According to an aspect of the present disclosure, a biological sample may be placed in a second disposable cartridge sample welland secured with a second disposable cartridge sample cap.

1412 1412 1412 The second disposable cartridge cassette bodymay be placed on a reader (not shown) for calibration. Saline pinch valves (not shown) in the second disposable cartridge cassette bodymay be activated when the second disposable cartridge cassette bodyis placed on a reader for calibration.

1404 1402 1406 1403 1413 1408 1408 1409 1404 1406 1404 1406 1409 1412 The second disposable cartridge sample wellcontaining the sample secured with a second disposable cartridge sample capis attached to a saline wellto form a sample-saline complexand loaded onto the cassette top covervia a docking location. The docking locationcomprises at least two loading wells. In one aspect of this exemplary embodiment, the second disposable cartridge sample welland the saline wellcomprise foils securing the respective liquids in their respective well. When the second disposable cartridge sample welland the saline wellare attached to the loading wells, the foils are pierced and the saline pinch valve is opened, exposing saline and the sample to channels (not shown) in the second disposable cartridge cassette body.

1410 1412 1410 1410 1400 According to an aspect of the present disclosure, the saline pinch valves are designed to be opened first to allow saline to flow over the SAW biosensor and contact the absorbent wicking padon the second disposable cartridge cassette body. In an illustrative embodiment, the absorbent wicking padpulls the saline at a rate of 5ul/min. Proceeding the opening the saline pinch valve for 5 minutes, the saline pinch valve is closed and the sample is allowed to be exposed to the SAW biosensor and then contact the absorbent wicking pad. The SAW biosensor may be in direct communication with the printed circuit board which in turn processes the output of the SAW biosensor due to the biological sample and an output of measurement (e.g., presence of an infectious virus in the biological sample) may be noted by the reader when the second disposable cartridge systemis placed in the reader.

15 15 FIGS.A andB 1500 1500 1515 1508 100 1518 600 100 1518 1518 1512 1519 1517 1518 1511 1506 1515 1500 1502 1503 1504 1515 1508 1502 1509 1506 1512 100 1508 1510 100 1518 100 1500 present a perspective and exploded view of a further exemplary embodiment of third disposable cartridge systemcomprising a SAW biosensor for the detection of infectious diseases, chemicals, proteins, nucleotides and cellular elements. The exemplary embodiment of the disposable cartridge systemcomprises a third disposable cartridge cassette bodyfurther comprising a saline blister pack, SAW biosensorattached to a printed circuit board, and a polymer gasketto separate the biological sample from coming in contact with the SAW biosensorand the printed circuit board. The printed circuit boardis secured onto the third disposable cartridge cassette bodyvia vertical postswhich fit on the post voidson the perimeter of the printed circuit board. The biological sample may be placed in a sample welland secured with a flexible sample capon the third disposable cartridge cassette body. The disposable cartridge systemmay be secured onto a reader (not shown) for calibration. Following calibration, a reusable flexible hourglass with grainsheld by a holding platformof a timer holdermay be placed on the third disposable cartridge cassette body, leading to opening the saline blister pack. As grains in the reusable flexible hourglass with grainsslowly transfer to the bottom of the hourglass and compress the indented regionof a flexible sample cap, the biological sample may be forced into the channel (not shown in the figures) in the third disposable cartridge cassette body, and allowed to interact with the SAW biosensorEventually the saline from the saline blister packand the biological sample are displaced to the waste well with an air vent. The SAW biosensormay be in direct communication with the printed circuit boardwhich in turn processes the output of the SAW biosensordue to the biological sample and an output of measurement (e.g., presence of a virus in the biological sample) may be noted in the reader when the disposable cartridge systemis placed in the reader. Also included could be a vibration module to shake the liquid contained in the flow cell.

16 FIG. 1600 1600 1602 1606 1618 1609 1608 1602 1616 1614 1604 1604 1609 1608 1606 1610 1612 Referring now to, a perspective view of an exemplary embodiment of a SAW biosensor cartridge systemadapted for collection and detection of gaseous species from the atmosphere is presented. The SAW biosensor cartridge systemincludes a gaseous species cassettewith a gaseous species sensorwhich may include a liquid-proof sealing, only revealing a sensing areacoated with an active nanomaterial layer. The gaseous species cassetteincludes a porous inletwith a filterfor gaseous species input from the atmosphere. The gaseous species input may travel through a gaseous species channelconnected to a micropump (not shown in the figure) allowing the gaseous species input to flow through the gaseous species channelfor interaction with the sensing areacoated with an active nanomaterial layer. In one embodiment, the data from the gaseous species sensormay then be transmitted to a reader via wireless radio frequency interrogationconnected to a tablet.

17 FIG. 22 FIG. 1700 1706 1710 1618 1716 1714 1706 1722 1720 1720 1712 1718 1712 1712 1716 1714 1710 1610 1612 Referring now to, a perspective view of an exemplary embodiment of a SAW biosensor cartridge system adapted for collection and detection of bioaerosols from the atmosphere. The second SAW biosensor cartridge systemincludes a bioaerosol cassettewith a bioaerosol sensorwhich may include a liquid-proof sealing, only revealing a bioaerosol sensing areacoated with a capture agent. The bioaerosol cassetteincludes a bioaerosol sampler systemwith a bioaerosol collection reservoirfor holding the bioaerosol input from the atmosphere. The bioaerosol collection reservoiris fluidly connected to a bioaerosol fluidic channelvia a valve. The sample bioaerosol may travel through the bioaerosol fluidic channelconnected to a micropump (not shown in the figure) allowing the bioaerosol input to flow through the bioaerosol fluidic channelfor interaction with the bioaerosol sensing areacoated with the capture agent. In one embodiment, the data from the bioaerosol sensormay then be transmitted to a reader via wireless radio frequency interrogationconnected to a tablet.represents capture of chemical and infectious disease agents from waste water or ground water or potable water into the flow cell with or without concentration of said sample.

Aspects of the present disclosure further includes a disposable cartridge system utilizing a surface acoustic wave (SAW) biosensor, disposable PCB assembly, fluid gasket, buffer can that contains saline, and a screw cap that can be used for the detection of infectious agents. A vibration module can also be added to shake up the reaction mix.

18 FIG.A 18 FIG.B 18 18 FIGS.A andB 18 FIG.B 1810 1810 1810 1810 1812 1814 1814 1814 1816 1810 1818 1822 1814 1818 1810 1814 1818 1814 1820 1810 1824 1826 shows a perspective view of the screw cap disposable cartridge, andpresents an exploded view of the screw cap disposable cartridge. According to some embodiments, screw cap disposable cartridgecan be docked onto a holder and wait for calibration. As shown in, screw cap disposable cartridgeincludes top foildisposed over buffer can, which can be removed by a user once calibration is complete to pierce a foil (not shown) at the bottom of buffer can. As shown in, buffer canis configured to mated with cylinderduring assembly. Screw cap disposable cartridgealso includes screw capand snap arm, which can be activated by a user to activate the flow of, for example, saline from the buffer canthrough the channelover the sensory aspect (not shown) of screw cap disposable cartridge. This permits the user to add the sample (not pictured) to the buffer canand turn the screw capagain, which activates the flow of the sample from the buffer canthrough the screw cap channelover the sensory aspect of screw cap disposable cartridge. According to the example embodiments, each of these components are located within perimeter, and is disposed on base.

19 FIG. 1900 1900 presents a data tabledemonstrating the ability of the systems of the present disclosure to detect SARS-COV-2 serology in clinical samples. According to the illustrative example, a sensor surface was immobilized with a recombinant SARS-COV-2 spike receptor-binding domain (RBD) protein for capturing SARS-COV-2 IgG antibodies. SARS-COV-2 IgG positive (n=10) and negative (n=10) human serum samples were evaluated without any sample processing with results obtained on average in 5.5 minutes. The results from this preliminary study indicate ability of the system to detect SARS-COV-2 IgG antibodies in clinical samples with 100% sensitivity and 100% specificity. Thus, data tableshows that the platform and the systems and devices of the present disclosure have the potential to be a POC method for improved detection of host generated antibodies against SARS-COV-2 and other infectious diseases.

20 FIG. presents graphs and data tables demonstrating the ability of gold coated surfaces to detect Canine Heart Worm Antigen (CHW) and canine Anti-Mullerian Hormone (AMH) via ELISA. According to the illustrative embodiment, the gold coated surfaces were immobilized with antibody-based capture agents for detecting CHW and canine AMH, respectively. The data demonstrates quantitative detection at picogram levels, which can correlate directly to sensor function.

21 FIG. Borrelia burgdorferi Referring now to, data demonstrating the ability of the systems of the present disclosure to detect host generated antibodies against(Bb) in clinical samples is presented. According to the illustrative embodiment, a sensor surface was immobilized with a recombinant antigen that consists of full-length DbpA, PepC10, C6 for capturing Bb specific IgM and IgG antibodies. Lyme Disease IgM positive (n=5), Lyme Disease IgG positive (n=7), and Lyme Disease IgM/IgG negative plasma samples were evaluated without any sample processing The results from this preliminary study indicate the ability of the system to detect Lyme Disease IgM/IgG antibodies in clinical samples with 100% sensitivity and 100% specificity. In addition, detection in whole blood was demonstrated without any sample processing. There was no significant change between plasma-spiked whole blood and plasma alone (P<0.05). The Lyme Disease assay results of the systems of the present disclosure were compared to a FDA/CE mark commercial IgM/IgG ELISA kit. The ELISA kit had 3 false positive and 3 equivocal results while the systems of the present disclosure had no false positive or false negative results. Thus, this data further demonstrates that the platform and the systems and devices of the present disclosure have the potential to be a POC method for improved detection of host generated antibodies against infectious diseases.

22 FIG. 2200 2200 2202 2206 1618 2208 1714 2202 2216 2204 2204 2208 1714 2206 1610 1612 presents a perspective view of an exemplary embodiment of a third SAW biosensor cartridge systemadapted for collection and detection of biological agents, such as infectious diseases, and chemical toxins, such as PFAS, from the waste water, potable water and ground water. The third SAW biosensor cartridge systemincludes a waste water cassettewith a waste water sensorwhich may include a liquid-proof sealing, only revealing a waste water sensing areacoated with a capture agent. The waste water cassetteincludes an inlet with a membrane filterto remove large particulates from the water sample input. The water sample input may travel through a waste water channelconnected to a micropump (not shown in the figure) allowing the water sample input to flow through the waste water channelfor interaction with the waste water sensing areacoated with the capture agent. In one embodiment, the data from the waste water sensormay then be transmitted to a reader via wireless radio frequency interrogationconnected to a tablet.

There are many instances when a rapid, portable, and accurate testing platform can add critical data to diagnose, leading to diagnosis of rapidly evolving biological events including proper treatment and decrease spread of infectious agents or detect the antibody reaction to such agents. Such a system, which can trace, analyze and handle large amounts of data related to such a diagnosis, is critical for prevention, treatment, and managing future outbreaks or follow disease course, therapeutic efficacy and safety and provide critical public health data in real time basis. Other examples of such utility can include non-infectious conditions such as chemo or bio toxin threats, where remote and constant monitoring can identify and isolate a person or non-persons carrying or distributing such materials using this technology. Other examples include measure of biomarkers to detect illness or wellness and oncologic markers on cell surface or in liquid phase to detect such markers. In addition, diagnosis of veterinary and human situations (disease and wellness) that can identify and treat rapidly can make a difference such as biomarkers which can rapidly identify traumatic brain injury, stroke, or myocardial infarction. Course of disease and therapy can also be monitored using these biomarkers and our system.

Although aspects of the present disclosure and certain examples are described herein in terms of biological, waste water and ground water samples or infectious samples, it should be understood by persons having ordinary skill in the art that the aspects and various alternative embodiments of the present disclosure could be performed on or implemented to detect non-biological and/or non-infectious analytes or samples and the like, within the scope of the present disclosure.

Although various aspects of the present disclosure are described herein in terms of various exemplary embodiments, it should be understood that variations and modifications may be made to the disclosure described herein to adopt it to various usages and conditions within the scope of applicant's invention as claimed herein.

The recitation of a listing of elements in any definition of a variable herein includes definitions of that variable as any single element or combination (or sub-combination) of listed elements. The recitation of an embodiment herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof.

All patents and publications mentioned in this specification are herein incorporated by reference to the same extent as if each independent patent and publication was specifically and individually indicated to be incorporated by reference.